Plant DNA extraction and purification integrated equipment

Abstract

The utility model relates to the technical field of DNA extraction and purification equipment, in particular to plant DNA extraction and purification integrated equipment which comprises a bottom plate, fixed vertical plates are arranged on the left side and the right side of the top of the bottom plate, a shaking assembly is movably connected between the two fixed vertical plates, the shaking assembly comprises a frame-shaped support, and a rectangular container is supported by the frame-shaped support. The top of the rectangular container is sleeved with a cover, a magnetic bead is placed in the rectangular container, a mounting plate is arranged between the two fixed vertical plates and close to the rear side of the rectangular container, and an electromagnet is arranged on the front side of the mounting plate. According to the plant DNA extraction and purification integrated equipment, the electromagnet accurately controls generation and disappearance of a magnetic field in a power-on and power-off mode, so that magnetic beads can be efficiently adsorbed or released at different stages, the problem that in the background technology, the magnetic beads are difficult to completely release after being adsorbed by the magnet is solved, full utilization of the magnetic beads is achieved, and the utilization rate of the plant DNA is increased. Meanwhile, the DNA extraction and purification efficiency and yield are improved.

Description

TECHNICAL FIELD

[0001] The utility model relates to DNA extraction and purification equipment technical field, concretely is plant DNA extraction and purification integrated equipment. BACKGROUND

[0002] Plant DNA extraction and purification is a key step in molecular biology research, and is widely used in genomics analysis, crop improvement, genetic diversity research and other fields. The purpose of extraction and purification is to separate high-quality genomic DNA from plant samples (such as leaves, seeds, roots, etc.) for subsequent PCR amplification, gene sequencing and other experiments. Magnetic bead method is one of the mainstream technologies for DNA extraction and purification. The magnetic bead method combines DNA through the special coating on the surface of the magnetic bead, and realizes rapid separation under the action of external magnetic field. Compared with traditional phenol-chloroform extraction method and centrifugal sedimentation method, the magnetic bead method has the advantages of simple operation, safety and environmental protection, high purification efficiency, etc. Its core process includes sample lysis, magnetic bead binding DNA, magnetic field separation DNA, impurity removal by washing, and finally DNA elution by buffer.

[0003] The patent with publication number CN208414401U discloses a one-stop magnetic bead extraction and purification DNA device, which comprises a placing cup, a rotating rod is arranged inside the placing cup, a support rod is arranged on the surface of the rotating rod, a stirring plate is arranged at one end of the support rod, a leakage groove is arranged on the surface of the placing cup, a cover plate is arranged on the top of the placing cup, a feeding port is arranged on the surface of the cover plate, a spring is arranged inside the cover plate, a clamping block is arranged on one side of the spring, and a cutout is arranged on the top of the leakage groove. The utility model discloses a leakage groove, which is conducive to taking out the biological magnetic beads combined with DNA. By rotating the holding sleeve to drive the rotating rod to rotate, the rotating rod drives the stirring plate to rotate through the support rod, so that the cell liquid is quickly contacted with the biological magnetic beads, and then the two are combined. The magnetic force of the magnet makes the biological magnetic beads with DNA closely adhere to the inner wall of the placing cup. The sliding block and the magnet are moved upward by the surrounding ring. The sliding block slides in the sliding groove, and then the magnet drives the biological magnetic beads to move.

[0004] The prior art has the following defects: the magnetic force of the magnet makes the biological magnetic beads with DNA closely adhere to the inner wall of the placing cup, the sliding block slides in the sliding groove, and then the magnet drives the biological magnetic beads to move, when moving to the leakage groove, the biological magnetic beads are adhered to the surface of the magnet through the leakage groove, then the ring is loosened, under the action of gravity, the magnet falls into the liquid storage groove, then the magnet is continuously lifted, then the liquid on the inner wall of the liquid storage groove is poured out, after being eluted by the elution solution, relatively purified DNA is obtained, but the magnet can only adsorb limited magnetic beads at a time, and after the magnetic beads are adsorbed by the magnet and then moved to the liquid storage groove, the magnetic beads are still adsorbed on the magnet, so that the magnet cannot be lifted again to adsorb the remaining magnetic beads in the placing cup, and the magnetic beads closely adhered to the surface of the magnet cannot be completely released into the elution solution for sufficient elution, so that the DNA recovery is incomplete, the yield of extracted DNA is limited, and defects exist in use. Content of the utility model

[0005] The utility model discloses a plant DNA extraction and purification integrated equipment, which aims at solving the problems in the prior art.

[0006] To achieve the above-mentioned purpose, the utility model provides the following technical scheme:

[0007] The plant DNA extraction and purification integrated equipment comprises a bottom plate for providing stability and bearing capacity of the whole device, left and right sides of the top of the bottom plate are each provided with a fixed vertical plate for providing support and fixing functions, a left-right movable shaking assembly is movably connected between the two fixed vertical plates, the shaking assembly comprises a frame-shaped support capable of sliding left and right, is used for driving the frame-shaped support and the rectangular container to move, so as to realize the mixing or uniform treatment of samples and magnetic beads, the frame-shaped support supports a rectangular container, and the rectangular container is a main container for sample treatment and is used for containing reagents such as magnetic beads, lysis solution, washing solution and elution solution.

[0008] The bottom of the rectangular container is of a conical structure, so that liquid can flow to the liquid outlet pipe at the bottom, the bottom end of the rectangular container is provided with a liquid outlet pipe with a valve, which is used for controlling the discharge of liquid, and the liquid can be discharged through the liquid outlet pipe during the separation or cleaning of the magnetic beads, so as to facilitate the multi-step treatment of samples, a lid is sleeved on the top of the rectangular container and is used for closing the container to prevent liquid leakage, and the rectangular container contains magnetic beads, which are key materials for extracting and purifying DNA, the magnetic beads are combined with DNA through a special coating on the surface of the magnetic beads, and the magnetic beads can realize separation, cleaning and elution under the action of an electromagnet.

[0009] Two said fixed vertical plate between and close to the position of the rear side of the rectangular container is equipped with mounting plate, the front side of the mounting plate is equipped with electromagnet, electromagnet outside power supply and controller work, for the rectangular container in the magnetic beads exert magnetic field, electromagnet through on-off control magnetic field generation or disappear, realize in different stage to the adsorption or release of magnetic beads, electromagnet and rectangular container does not contact, avoid the rectangular container when moving left and right wear and tear.

[0010] Preferably, the top of the bottom plate and near the four corners are provided with mounting holes, which can be fixed on the workbench by bolts as needed, and the bottom of the bottom plate is attached with a non-slip pad with a matching shape and size, which can increase the stability of the equipment during operation and prevent sliding or tilting.

[0011] Preferably, the middle part of the left and right sides of the frame-shaped support is provided with a rectangular movable rod, and the side surface of the fixed vertical plate is provided with a rectangular hole for the rectangular movable rod to pass through, so as to allow the rectangular movable rod to pass through and provide sliding space for the movement of the shaking assembly, ensuring that the shaking assembly can move left and right.

[0012] Preferably, the outside of the rectangular movable rod is sleeved with a spring for providing a rebounding force, one end of the spring abuts against the side surface of the frame-shaped support, and the other end of the spring abuts against the side surface of the fixed vertical plate. When the shaking assembly moves, the spring is compressed, and when it is released, it pushes back, so that the shaking assembly drives the rectangular container to shake, facilitating the full contact of the solution with the magnetic beads.

[0013] Preferably, the inner wall of the frame-shaped support is provided with a rubber lining, and the inner side of the rubber lining is tightly attached to the outer side of the rectangular container. The function of the rubber lining is to fix the rectangular container and play a buffering and damping role, preventing the container from being damaged or loosened due to shaking.

[0014] Preferably, the middle and upper part of the outer side of the rectangular container is provided with a frame-shaped limiting plate, and the bottom of the frame-shaped limiting plate abuts against the top of the frame-shaped support. The frame-shaped limiting plate is used to further stabilize the rectangular container and prevent it from moving downward due to shaking, so that the rectangular container remains in a hovering state.

[0015] Preferably, the side surface of the inner wall of the cover is provided with a frame-shaped rubber, and the inner side of the frame-shaped rubber is tightly attached to the outer side of the rectangular container, which is used to enhance the sealing performance of the container and prevent liquid from leaking out of the top of the container.

[0016] Preferably, the top of the cover is provided with a liquid injection pipe for injecting reagent or sample liquid into the rectangular container, and the top of the liquid injection pipe is hinged with a flip cover for closing the liquid injection port to prevent impurities from entering.

[0017] Compared with the prior art, the beneficial effects of the present application are:

[0018] 1. The plant DNA extraction and purification integrated device, the electromagnet precisely controls the generation and disappearance of the magnetic field through on-off electricity, so that the magnetic beads can be efficiently adsorbed or released at different stages, avoiding the problem that the magnetic beads are difficult to completely release after being adsorbed by the magnet in the background technology, thereby realizing the full utilization of the magnetic beads and improving the efficiency and yield of DNA extraction and purification.

[0019] 2. The plant DNA extraction and purification integrated device, through the design of the shaking assembly, the rectangular container can shake left and right, and the reset force of the spring ensures that the magnetic beads are fully mixed in the lysis solution, the washing solution and the eluent. This design overcomes the problem of uneven contact between the magnetic beads and the liquid in the background technology, significantly improves the binding efficiency of DNA and magnetic beads and the elution effect, thereby ensuring the recovery rate and purity of the target DNA.

[0020] 3. The plant DNA extraction and purification integrated device, through the non-contact design of the electromagnet and the rectangular container, the magnetic beads are prevented from directly contacting the electromagnet, and in addition, after the electromagnet is powered off, the magnetic beads can quickly escape from the magnetic field and fully contact the eluent, ensuring that the DNA can be completely released into the eluent, greatly improving the integrity of DNA purification. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 It is a whole first visual angle structure schematic view of the utility model;

[0022] Figure 2 It is a whole second visual angle structure schematic view of the utility model;

[0023] Figure 3 It is a partial structure schematic view of the utility model;

[0024] Figure 4 It is an assembly structure schematic view of the fixed vertical plate, the shaking assembly and the rectangular container in the utility model;

[0025] Figure 5 It is a rectangular container structure schematic view in the utility model;

[0026] In the figure: 1, bottom plate; 2, fixed vertical plate; 20, rectangular hole; 3, shaking assembly; 30, frame-shaped support; 31, rectangular movable rod; 32, spring; 33, rubber lining; 4, rectangular container; 40, liquid outlet pipe; 41, cover; 410, liquid injection pipe; 411, flip cover; 42, frame-shaped rubber; 43, frame-shaped limiting plate; 5, mounting plate; 6, electromagnet; 7, magnetic beads; 8, non-slip pad. DETAILED DESCRIPTION

[0027] Clearly and completely describe the technical scheme in the embodiments of the utility model with reference to the drawings in the embodiments of the utility model, obviously, the described embodiments are only some embodiments of the utility model, and not all embodiments. Based on the embodiments in the utility model, all other embodiments obtained by the ordinary skilled in the art without creative labor belong to the scope of the utility model protection.

[0028] In the description of the utility model, it is understood that the orientation or position relationship indicated by the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise" and the like is based on the orientation or position relationship shown in the drawings, and is only for the convenience of describing the utility model and simplifying the description, and does not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the utility model.

[0029] Please refer to Figures 1-5 The utility model provides a technical scheme:

[0030] The plant DNA extraction and purification integrated equipment, including bottom plate 1, be used for providing the stability and carrying capacity of overall device, the left and right sides of the top of bottom plate 1 are equipped with fixed vertical plate 2, be used for providing support and fixed function, the movable connection of two fixed vertical plate 2 has the left and right movable shaking assembly 3, shaking assembly 3 includes frame support 30, can left and right slide, be used for driving frame support 30 and rectangular container 4 move, to realize sample and magnetic bead 7 mixing or uniform processing, frame support 30 supports rectangular container 4 rectangular container 4 is the main container of sample processing, is used for accommodating magnetic bead 7, lysate, washing liquid and eluent and other reagents;

[0031] The bottom of rectangular container 4 is conical structure, facilitates the flow of liquid to the bottom outlet pipe 40, the bottom end of rectangular container 4 is equipped with the outlet pipe 40 with valve, is used for controlling the discharge of liquid, in the process of magnetic bead 7 separation or cleaning, liquid is discharged through outlet pipe 40, the multi-step processing of sample is convenient, the top of rectangular container 4 is equipped with lid 41, is used for closing container, prevents liquid leakage, rectangular container 4 is placed with magnetic bead 7, is the key material of extraction and purification DNA, magnetic bead 7 is combined with DNA through the special coating on its surface, and realizes separation, cleaning and elution and other functions under the action of electromagnet 6;

[0032] An installation plate 5 is arranged between the two fixed vertical plates 2 and close to the rear side of the rectangular container 4. The front side of the installation plate 5 is provided with an electromagnet 6. The electromagnet 6 is connected with a power supply and a controller and is used to apply a magnetic field to the magnetic beads 7 in the rectangular container 4. The electromagnet 6 controls the generation or disappearance of the magnetic field by switching on or off, so as to realize the adsorption or release of the magnetic beads 7 in different stages. The electromagnet 6 does not contact the rectangular container 4, thereby avoiding abrasion of the rectangular container 4 when moving left and right.

[0033] In the embodiment, mounting holes are arranged at the top of the bottom plate 1 and close to the four corners. The bottom plate 1 can be fixed on the workbench by bolts as needed. The bottom of the bottom plate 1 is attached with a non-slip pad 8 with a shape and size matching the bottom plate 1. The non-slip pad 8 can increase the stability of the device during operation and prevent sliding or tilting.

[0034] Specifically, the middle part of the left and right sides of the frame-shaped support 30 is provided with a rectangular movable rod 31. The side surface of the fixed vertical plate 2 is provided with a rectangular hole 20 for the rectangular movable rod 31 to pass through, so as to allow the rectangular movable rod 31 to pass through and provide a sliding space for the movement of the shaking assembly 3, thereby ensuring that the shaking assembly 3 can move left and right.

[0036] Further, the outer side of the rectangular movable rod 31 is sleeved with a spring 32 for providing a rebounding force. One end of the spring 32 abuts against the side surface of the frame-shaped support 30, and the other end of the spring 32 abuts against the side surface of the fixed vertical plate 2. When the shaking assembly 3 moves, the spring 32 is compressed and releases the rebounding force when released, so as to drive the rectangular container 4 to shake, which is conducive to the full contact between the solution and the magnetic beads 7.

[0037] Further, the inner wall of the frame-shaped support 30 is provided with a rubber lining 33. The inner side of the rubber lining 33 tightly abuts against the outer side of the rectangular container 4. The function of the rubber lining 33 is to fix the rectangular container 4 and play a buffering and damping role, thereby preventing the container from being damaged or loosened due to shaking.

[0038] Further, the middle upper part of the outer side of the rectangular container 4 is provided with a frame-shaped limiting plate 43. The bottom of the frame-shaped limiting plate 43 abuts against the top of the frame-shaped support 30. The frame-shaped limiting plate 43 is used to further stabilize the rectangular container 4 and prevent it from moving downward due to shaking, so as to keep the rectangular container 4 in a hovering state.

[0039] Further, the side surface of the inner wall of the cover 41 is provided with a frame-shaped rubber 42. The inner side of the frame-shaped rubber 42 tightly abuts against the outer side of the rectangular container 4, which is used to enhance the sealing performance of the container and prevent liquid from leaking out of the top of the container.

[0040] Further, the top of the cover 41 is provided with a liquid injection pipe 410 for injecting reagent or sample liquid into the rectangular container 4. The top of the liquid injection pipe 410 is hinged with a flip cover 411 for closing the liquid injection port and preventing impurities from entering.

[0040] It should be noted that the specific operation steps of the integrated device are as follows:

[0041] Step 1: First, insert the rectangular container 4 into the frame-shaped support 30 until the bottom of the frame-shaped limiting plate 43 abuts against the top of the frame-shaped support 30, then place a plurality of magnetic beads 7 in the rectangular container 4, and then pour a buffer solution into the rectangular container 4, the buffer solution containing PEG and salt ions;

[0042] Step 2: Add a DNA sample, then cover the lid 41 on the top of the rectangular container 4, and then shake the frame-shaped support 30 left and right through the rectangular movable rod 31, so that the frame-shaped support 30 drives the rectangular container 4 to move left and right, so that the magnetic beads 7 in the rectangular container 4 are in full contact with the solution;

[0043] Step 3: Turn on the electromagnet 6 to adsorb the magnetic beads 7 to the inner wall of the rectangular container 4, so that the magnetic beads 7 and the liquid are separated, then the worker takes the idle container to take the waste liquid, places the idle container directly below the liquid outlet pipe 40, and opens the valve of the liquid outlet pipe 40, and then the liquid in the rectangular container 4 is completely discharged, after the solution is completely discharged, the valve of the liquid outlet pipe 40 is closed;

[0044] Step 4: Then the worker opens the flip cover 411, and adds ethanol cleaning to the rectangular container 4 through the liquid injection pipe 410, and adds 80% ethanol to clean the PEG and excess salt ions;

[0045] Step 5: After the cleaning operation is completed, open the valve of the liquid outlet pipe 40 to completely discharge the liquid generated by the ethanol cleaning, and then close the valve of the liquid outlet pipe 40, open the flip cover 411, and add an eluent to the rectangular container 4 through the liquid injection pipe 410, the eluent being water or TE Buffer resuspension, at this time, the electromagnet 6 is powered off, and the frame-shaped support 30 is shaken left and right, so that the magnetic beads 7 are in full contact with the eluent, and the DNA is eluted from the magnetic beads;

[0046] Step 6: After the elution operation is completed, turn on the electromagnet 6 to adsorb the magnetic beads 7 to the inner wall of the container, so that the magnetic beads 7 and the liquid are separated, then the worker takes the container for taking the supernatant and places it directly below the liquid outlet pipe 40, and opens the valve of the liquid outlet pipe 40, and then the supernatant in the rectangular container 4 is discharged.

[0047] In use, first, the rectangular container 4 is inserted into the frame-shaped support 30 until the bottom of the frame-shaped limiting plate 43 abuts against the top of the frame-shaped support 30, ensuring that the rectangular container 4 is securely installed, then the magnetic beads 7 are placed in the rectangular container 4, and then buffer solution (containing PEG and salt ions) is added into the rectangular container 4 through the liquid injection pipe 410, ensuring that the liquid covers the magnetic beads 7; then, the processed DNA sample is added into the rectangular container 4, the lid 41 is covered, and the frame-shaped rubber 42 on the inner side of the lid 41 is ensured to tightly adhere to the outer side of the rectangular container 4, preventing liquid leakage or pollution, the frame-shaped support 30 is shaken through the rectangular movable rod 31, the rectangular container 4 is moved left and right, ensuring that the magnetic beads 7 are in full contact with the sample liquid, and DNA can be efficiently combined to the surface of the magnetic beads 7; subsequently, the electromagnet 6 is powered on, the magnetic beads 7 are adsorbed to the inner wall of the rectangular container 4 and separated from the liquid, the valve of the liquid outlet pipe 40 is opened, the waste liquid in the container is discharged, the valve of the liquid outlet pipe 40 is closed, the flip cover 411 is opened, 80% ethanol solution is added into the rectangular container 4 through the liquid injection pipe 410, the PEG and excess salt ions on the surface of the magnetic beads 7 are cleaned, the frame-shaped support 30 is shaken again through the rectangular movable rod 31, ensuring that the magnetic beads 7 are in full contact with the ethanol, then the valve of the liquid outlet pipe 40 is opened, and the cleaning liquid is discharged; subsequently, the valve of the liquid outlet pipe 40 is closed, the flip cover 411 is opened, elution liquid (such as water or TE buffer solution) is added into the rectangular container 4 through the liquid injection pipe 410, the power supply of the electromagnet 6 is disconnected, the magnetic beads 7 are released from the inner wall of the container, the frame-shaped support 30 is shaken again through the rectangular movable rod 31, the magnetic beads 7 are in full contact with the elution liquid, and DNA is eluted from the surface of the magnetic beads 7; finally, the electromagnet 6 is powered on, the magnetic beads 7 are adsorbed to the inner wall of the rectangular container 4 and separated from the liquid, the valve of the liquid outlet pipe 40 is opened, the supernatant containing DNA is discharged and collected into a special container, and the DNA extraction and purification operation is completed.

[0048] The basic principle, main features and advantages of the present application are shown and described above. It should be understood by those skilled in the art that the present application is not limited by the above embodiments, and the above embodiments and descriptions in the specification are only preferred examples of the present application and are not intended to limit the present application. Without departing from the spirit and scope of the present application, various changes and improvements can be made to the present application, and these changes and improvements all fall within the scope of the claimed present application. The scope of protection of the present application is defined by the appended claims and their equivalents.

Claims

1. A plant DNA extraction and purification integrated device, comprising a base plate (1), characterized in that: The left and right sides of the top of the bottom plate (1) are provided with fixed vertical plates (2), and the movable shaking assembly (3) is movably connected between the two fixed vertical plates (2), the shaking assembly (3) comprises a frame-shaped support (30), the frame-shaped support (30) supports a rectangular container (4), the bottom of the rectangular container (4) is of a conical structure, the bottom end of the rectangular container (4) is provided with a liquid outlet pipe (40) with a valve, the top of the rectangular container (4) is provided with a cover (41), the rectangular container (4) is placed with magnetic beads (7), the position between the two fixed vertical plates (2) and close to the rear side of the rectangular container (4) is provided with a mounting plate (5), and the front side of the mounting plate (5) is provided with an electromagnet (6).

2. The plant DNA extraction and purification integrated device according to claim 1, characterized in that: The top of the bottom plate (1) and the positions close to the four corners are provided with mounting holes, and the bottom of the bottom plate (1) is attached with a non-slip pad (8) with a shape and size matching.

3. The plant DNA extraction and purification integrated device according to claim 1, characterized in that: The middle of the left and right sides of the frame-shaped support (30) is provided with a rectangular movable rod (31), and the side surface of the fixed vertical plate (2) is provided with a rectangular hole (20) for the rectangular movable rod (31) to pass through.

4. The plant DNA extraction and purification integrated device according to claim 3, characterized in that: The outer side of the rectangular movable rod (31) is provided with a spring (32), one end of the spring (32) abuts against the side surface of the frame-shaped support (30), and the other end of the spring (32) abuts against the side surface of the fixed vertical plate (2).

5. The plant DNA extraction and purification integrated device according to claim 1, characterized in that: The inner wall of the frame-shaped support (30) is provided with a rubber lining (33), and the inner side of the rubber lining (33) is tightly attached to the outer side of the rectangular container (4).

6. The plant DNA extraction and purification integrated device according to claim 1, characterized in that: The middle upper part of the outer side of the rectangular container (4) is provided with a frame-shaped limiting plate (43), and the bottom of the frame-shaped limiting plate (43) abuts against the top of the frame-shaped support (30).

7. The plant DNA extraction and purification integrated device according to claim 1, characterized in that: The side surface of the inner wall of the cover (41) is provided with a frame-shaped rubber (42), and the inner side of the frame-shaped rubber (42) is tightly attached to the outer side of the rectangular container (4).

8. The plant DNA extraction and purification integrated device according to claim 1, characterized in that: The top of the cover (41) is provided with a liquid injection pipe (410), and the top of the liquid injection pipe (410) is hinged with a flip cover (411).

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